What information is needed to begin a custom antibody production project?

To begin a new project, we require detailed information about your target antigen, including its sequence or structure, and the desired application for the final antibody. Providing details about the target protein's function, its known epitopes, and any cross-reactivity concerns helps our team to design the most effective strategy for immunogen design and screening. Any existing data on the antigen is helpful for project success.

How do you handle intellectual property (IP) and confidentiality for my project?

We take IP and confidentiality very seriously and have strict protocols in place to protect your information. All projects are conducted under a confidential disclosure agreement (CDA) before any proprietary details are shared. Any antibodies, cell lines, or other materials developed during the project are the sole property of the client upon completion and final delivery.

What QC tests are performed to ensure the antibody is suitable for IVD use?

We perform a comprehensive suite of QC tests to ensure your antibody meets IVD-grade standards. This includes verifying antibody concentration via UV spectrophotometry, assessing purity using SDS-PAGE and HPLC, and confirming binding specificity and affinity through ELISA and surface plasmon resonance (SPR). Additionally, we test for endotoxin levels and assess stability under various storage conditions.

Can you help me decide between developing a monoclonal or a polyclonal antibody?

Yes, our expert team provides consultation to help you make this crucial decision based on your specific application and project goals. MAbs offer high specificity and excellent batch-to-batch consistency, making them ideal for high-precision sandwich assays. Polyclonal antibodies provide broader epitope recognition and are often more robust for detecting low-abundance proteins, making them suitable for applications like immunohistochemistry.

What if the antibody you produce does not perform as expected in my assay?

Our goal is to ensure the successful performance of your custom antibody in its intended application. In the unlikely event that the antibody does not meet the agreed-upon performance specifications, our team will work closely with you to troubleshoot the issue. We will conduct a thorough investigation and, if necessary, re-initiate the production process at no additional cost to meet the project specifications.

How do you ensure lot-to-lot consistency for long-term production?

We ensure exceptional lot-to-lot consistency through a combination of using a single, validated hybridoma cell line for mAbs, following rigorous standard operating procedures (SOPs) for all production steps, and performing extensive QC testing on every production lot. Each batch is compared to a reference standard to guarantee functional equivalence and reliable performance over the long term.

Custom IVD Antibody Production Service

Antibodies are crucial in in vitro diagnostics (IVD) research and therapeutics, driving continuous demand. Creative Biolabs, with extensive expertise, supports IVD antibody development. Our advanced IVD antibody development platform facilitates the discovery and production of high-affinity, high-specificity polyclonal and monoclonal antibodies (mAbs). We employ diverse technologies like hybridoma, single B cell sorting, and various display systems. High-quality IVD antibodies are then produced using a comprehensive array of expression systems, including bacterial, yeast, and mammalian cells.

Expression Systems for IVD mAbs Production

Fig.1 Overview of IVD mAbs production. (Creative Biolabs Original)

Fig.2 Bacteria. (Creative Biolabs Authorized)

Bacteria expression system

Bacteria are very attractive for use in antibody production because of their fast growth, ease of manipulation, relatively well-known information and much lower antibody production costs. Production of recombinant antibody fragments in Escherichia coli has been focused mainly on two major formats of antibody fragments: single-chain variable fragment (scFv) and antigen-binding fragment (Fab). Creative Biolabs has established several systems for antibody fragment production, such as high expression systems with T7 RNA polymerase gene promoter, coexpression of foldases or molecular chaperones to prevent misfolding and aggregation of antibody fragments.

Yeast expression system

Although the pattern of N-glycan in yeast is quite different from the human counterparts, yeasts have their own glycosylation machinery. With further optimization of the host and the production process, yeast has great potential to become a viable platform to produce competitive yields of complex and functional antibodies. At Creative Biolabs, significant progress has been made in the engineering of yeast cells (Saccharomyces cerevisiae and Picchia pastoris) for enhanced production of IVD antibodies.

Fig.3 Yeast. (Creative Biolabs Authorized)

Fig.4 Mammalian cell. (Creative Biolabs Authorized)

Mammalian cell expression system

The most prominent host cell lines for antibody production are CHO, NS0, Sp2/0, HEK 293 and PER.C6. 1) CHO-based mAb production processes typically reach the highest product titers with 1 g/L (batch) and 1-10 g/L (fed-batch). 2) The most prominent characteristic of NS0 and Sp2/0 is that they express the two predominant immunogenic glycan epitopes for humans, galactose-alpha1,3-Gal (alpha-gal) and N-glycolyl-neuraminic acid (Neu5Gc). 3) HEK and PER.C6 cells are used to produce antibodies with human identical glycosylation. HEK293 cells are specially applied for transient gene expression and reach mAb expression levels only in the low tens to hundreds of milligrams per liter range during batch cultivation and up to 0.6 g/L (fed-batch). Regarding PER.C6 cells, product yields are more in the range of CHO-based processes with mAb titers of 0.5 g/L (batch), 8 g/L (fed-batch), and even 27 g/L (perfusion). Creative Biolabs offers mammalian cell transient expression and stable cell line development for the production of IVD mAbs with a volume that could reach 1000 L and even more.

Baculoviral insect cell expression system

Insect cells are a very powerful tool for the expression of large, highly processed antibodies because of their ability to perform signal-peptide cleavage, N- and O-linked glycosylation and efficient extracellular secretion. Under the control of very strong viral promoters, it is possible to transiently express large DNA fragments by using baculoviruses. What's more, the glycosylation patterns have been shown to differ somewhat as compared to mammalian cells. Spodoptera frugiperda (SF) and Trichoplusia ni are the main insect cell lines used for antibody production. Sf9, Sf21 and "High Five" cells are available at Creative Biolabs to support the production of IVD mAbs.

Fig.6 Plant. (Creative Biolabs Authorized)

Plant expression system

The plant as a bioreactor for the mAb production has several advantages, which include the lack of animal pathogenic contaminants, low cost of production, and ease of agricultural scale-up compared to other currently available systems. Thus, plants are considered to be a potential alternative to compete with other systems. Creative Biolabs has developed an advanced antibody expression platform based on Nicotiana benthamiana and N. tabacum to produce IVD antibodies.

Cell-free expression systems

Cell-free systems are emerging as an attractive alternative for the production of antibodies due to their flexible nature without any cell membrane constraints. Cell-free systems based on cell lysates derived from different sources, and with batch-to-batch consistency, have acted as a catalyst for the cell-free synthesis of target antibodies. Creative Biolabs provides several cell-free systems based on the cell lysate of prokaryotic or eukaryotic cells for the production of IVD antibodies.

Fig.7 Cell-free system. (Creative Biolabs Authorized)

Our Custom IVD Antibody Production Services

Creative Biolabs offers highly efficient antibody production services across a diverse array of expression systems. To facilitate the manufacture of IVD antibodies, Creative Biolabs has established specialized bacterial, yeast, mammalian cell, baculoviral insect cell, plant, and cell-free expression platforms. These systems enable comprehensive, one-stop services, encompassing project design, sequence optimization, gene synthesis, vector construction, and culminating in the production and purification of high-quality antibodies.

Our custom antibody production capabilities range from high-throughput, low-amount synthesis to large-scale production, with capacities up to 100 grams. Furthermore, our services include the production and purification of various antibody formats, such as recombinant IgG, scFv/Fab fragments, bispecific antibodies (BsAbs), and single domain antibodies (sdAbs).

Service Workflow of Custom IVD Antibody Production

01Project Design and Consultation

The process begins with an in-depth discussion to understand your specific project goals, target antigen characteristics, and the intended diagnostic application.

02Sequence Optimization and Gene Synthesis

We perform strategic optimization of antibody sequences to enhance expression and stability. This is followed by the precise synthesis of the gene sequences.

03Vector Construction

The synthesized genes are cloned into a suitable expression vector, preparing the genetic material for introduction into the host expression system.

04Expression System Selection and Antibody Production

Based on your project's needs, we select the most appropriate expression system to produce the antibody. This can involve recombinant expression in bacteria, yeast, mammalian cells, etc.

05Antibody Purification

Antibodies are harvested and purified using advanced chromatography techniques to achieve the desired level of purity and remove any contaminants.

06Quality Control (QC) and Validation

Every batch undergoes comprehensive QC analysis, including UV, SDS-PAGE, and ELISA, to confirm its purity, concentration, specificity, and stability.

07Final Delivery

We provide the finished, high-quality antibody product along with a detailed technical report and a certificate of analysis, ensuring a fully documented and traceable final delivery.

Applications

Immunoassay Development

Our antibodies are critical components for the creation of high-performance immunoassays, including ELISA, chemiluminescent immunoassay (CLIA), and lateral flow immunoassay (LFIA) platforms. We specialize in developing matched antibody pairs for enhanced sensitivity and specificity in sandwich assays.

Disease-Specific Biomarker Detection

We provide antibodies tailored for detecting specific biomarkers in clinical samples for various diseases. This includes diagnostics for infectious diseases (e.g., influenza, emerging pathogens), oncology (e.g., tumor markers), cardiovascular health, and fertility.

Cell and Tissue Analysis

Our antibodies are validated for use in cellular and tissue-based diagnostic methods such as immunohistochemistry (IHC), immunocytochemistry (ICC), and flow cytometry, allowing for precise visualization and quantification of target proteins in complex biological samples.

Published Data

1. Integration of Phage and Mammalian Cell Display Platforms for Germline-Like Monoclonal Antibody Production

Fig.1 Flow chart of the anti-TIM-3 mAbs selection strategy combining both phage and mammalian cell antibody display. Fig.1 The process of TIM-3-specific mAbs selection strategy.^1,3

In this work, researchers developed a novel platform combining phage and mammalian cell display to enable efficient selection of full-length IgG antibodies based on their binding affinity. Utilizing this method, researchers generated a potent germline-like anti-TIM-3 mAb, which demonstrated strong binding affinity, specificity, and therapeutic potential in vivo and in vitro studies. Therefore, this platform enables the efficient selection of full-length IgG antibodies with germline-like sequences and assists in developing high affinity and low immunogenicity mAbs for therapies.

2. A Rapid Cell-Free Expression and Screening System for Antibody Discovery

Fig.2 Schematic of the steps involved in the cell-free antibody screening workflow. Fig.2 Workflow of a rapid cell-free antibody screening.^2,3

This study presented an integrated workflow for antibody expression and evaluation, integrating cell-free DNA assembly and amplification, cell-free protein synthesis, and binding assessments of antibody fragments, enabling rapid screening within hours instead of weeks. The workflow was applied to assess 135 antibodies targeting SARS-CoV-2, including all 8 granted emergency use authorization for COVID-19, identifying the most potent candidates. Additionally, 119 antibodies derived from mice immunized against the SARS-CoV-2 spike glycoprotein were evaluated, revealing neutralizing antibodies, which exhibited potent binding and neutralization activity against a broad range of SARS-CoV-2 variants. This cell-free platform accelerates antibody discovery for future pandemics and broader research, diagnostic, and therapeutic applications.

Service Highlights

One-Stop Service: We provide a comprehensive, end-to-end service from initial project design and gene synthesis to high-quality antibody production and purification. This integrated approach simplifies your workflow.
Diverse Antibody Formats: We offer a wide selection of antibody formats to suit your application, including full-length antibodies (e.g., human IgGs) and specialized formats like scFv, Fab, and BsAbs.
Guaranteed High Purity and Low Endotoxin Levels: Our advanced purification protocols ensure antibodies achieve a purity of up to 99%. We also guarantee low endotoxin levels, which is a critical requirement for reliable in vitro diagnostic assays.
Comprehensive QC Analysis: We perform extensive QC testing at every stage, including UV spectrophotometry, SDS-PAGE, ELISA, and HPLC. This rigorous analysis ensures every batch meets the highest standards for specificity and stability.
Powered by Proven, First-in-Class Technology: Our services are built on industry-leading technologies and validated platforms. We continuously invest in advanced methods like hybridoma and display technologies to provide you with cutting-edge solutions.
Dedicated After-Sale Support: Our commitment extends beyond final delivery with dedicated after-sale support. We address technical questions and provide guidance to ensure your custom antibodies perform optimally in your assay.

FAQs

Q: What information is needed to begin a custom antibody production project?

A: To begin a new project, we require detailed information about your target antigen, including its sequence or structure, and the desired application for the final antibody. Providing details about the target protein's function, its known epitopes, and any cross-reactivity concerns helps our team to design the most effective strategy for immunogen design and screening. Any existing data on the antigen is helpful for project success.
Q: How do you handle intellectual property (IP) and confidentiality for my project?

A: We take IP and confidentiality very seriously and have strict protocols in place to protect your information. All projects are conducted under a confidential disclosure agreement (CDA) before any proprietary details are shared. Any antibodies, cell lines, or other materials developed during the project are the sole property of the client upon completion and final delivery.
Q: What QC tests are performed to ensure the antibody is suitable for IVD use?

A: We perform a comprehensive suite of QC tests to ensure your antibody meets IVD-grade standards. This includes verifying antibody concentration via UV spectrophotometry, assessing purity using SDS-PAGE and HPLC, and confirming binding specificity and affinity through ELISA and surface plasmon resonance (SPR). Additionally, we test for endotoxin levels and assess stability under various storage conditions.
Q: Can you help me decide between developing a monoclonal or a polyclonal antibody?

A: Yes, our expert team provides consultation to help you make this crucial decision based on your specific application and project goals. MAbs offer high specificity and excellent batch-to-batch consistency, making them ideal for high-precision sandwich assays. Polyclonal antibodies provide broader epitope recognition and are often more robust for detecting low-abundance proteins, making them suitable for applications like immunohistochemistry.
Q: What if the antibody you produce does not perform as expected in my assay?

A: Our goal is to ensure the successful performance of your custom antibody in its intended application. In the unlikely event that the antibody does not meet the agreed-upon performance specifications, our team will work closely with you to troubleshoot the issue. We will conduct a thorough investigation and, if necessary, re-initiate the production process at no additional cost to meet the project specifications.
Q: How do you ensure lot-to-lot consistency for long-term production?

A: We ensure exceptional lot-to-lot consistency through a combination of using a single, validated hybridoma cell line for mAbs, following rigorous standard operating procedures (SOPs) for all production steps, and performing extensive QC testing on every production lot. Each batch is compared to a reference standard to guarantee functional equivalence and reliable performance over the long term.

IVD research increasingly relies on mAbs. Creative Biolabs is pleased to present world-leading IVD antibody production services. For more detailed information, please feel free to contact us for a detailed quote and further discussion with our scientists.

References

Jin, Yu-jia, et al. "A novel and effective approach to generate germline-like monoclonal antibodies by integration of phage and mammalian cell display platforms." Acta Pharmacologica Sinica 43.4 (2022): 954-962.
Hunt, Andrew C., et al. "A rapid cell-free expression and screening platform for antibody discovery." Nature communications 14.1 (2023): 3897.
Distributed under Open Access license CC BY 4.0. The image was modified by extracting and using only part of the original image.